Co-enzymes and cofactors with an adenine ring system like AMP, ADP, ATP, NAD.sup.+ /NADH and NADP.sup.+ /NADPH, which are covalently bound to solid carriers or to soluble polymers, e.g. water soluble polymers, have already been widely used and examined. For example, co-enzymes bound to solid carriers are successfully used as ligands in affinity chromatography. Water soluble polymers with covalently bound co-enzymes are very useful in affinity partitioning. In the case of NAD.sup.+ /NADH and NADP.sup.+ /NADPH corresponding compounds can be regenerated and correspondingly used in enzymatical systems with recycling.
When producing co-enzymes covalently bound to macro-molecules and containing an adenine ring system, as is well known, a functional group is first provided at the N-atom in position 6 after which this reactive group is reacted with a macro-molecule.
First of all the adenine ring system is reacted at the N(1)-atom with an alkylating agent bearing an additional functional group which is designed to permit the reaction with the macro-molecule or a chain extension before the reaction with the macro-molecule. Alkylating agents include, for example, halo acids like iodine acetic acid, epoxides like 3.4-epoxy butyric acid, lactones, like propiolactone, or aziridines, like ethylene imine.
In the case of AMP, ADP and ATP the alkylation is immediately succeeded by a Dimroth rearrangement into the N.sup.6 -form. The Dimroth rearrangement can be immediately followed by the reaction with the macro-molecule or can be followed by the said reaction after a preceding chain extension at the N-atom at position 6. In the case of NAD.sup.+ and NADP.sup.+ a reduction is carried out before the Dimroth rearrangement and a reoxidation after the Dimroth rearrangement.
The co-enzyme derivatives produced in the same manner can bear, for example, the following functional groups on chains at the N-atom at position 6 for a linking with macro-molecules: carboxylic, amino, or vinyl groups.
For a linking insoluble macro-molecules (carrier, matrices) or soluble, especially water soluble, macro-molecules can be taken into consideration which have one or several functional groups effective for the linking. These macro-molecules may contain such functional groups per se or the said functional groups may be introduced into the macro-molecules, for example according to the bromic cyanide method. (Experts can obtain evidence of the possibility of linking the said macro-molecules by a simple experiment.) Examples for macro-molecules which can be used are: dextrans, polyetherpolyols like polyethyleneglycol, polyethylene-imines, polyacrylamides, co-polymers like poly-(methylvinylether/maleic acid anhydride), agarose, glass, cellulose and derivatives of the said macro-molecules; cf. Lowe, TIBS, 134 (1978) and Mosbach, Advances in Enzymology, 46 (1978) 205 to 278.
In the case of the known processes mentioned, the resulting yield after the Dimroth rearrangement (and after an optionally succeeding chain extension at the N-atom at position 6) has not so far proved satisfactory. The following Table lists yields in the range of merely 12 to 40% for (a) prior art.